Abrogation Of Gap Junctional Communication In Es Cells Results In A Disruption Of Primitive Endoderm Formation In Embryoid Bodies
Philipp Wörsdörfer, Felicitas Bosen, Martina Gebhardt, Nicole Russ, Katrin Zimmermann, David Komla Kessie, Thileepan Sekaran, Angela Egert, Süleyman Ergün, Hubert Schorle, Alexander Pfeifer, Frank Edenhofer and Klaus Willecke
Accepted manuscript online: 21 NOV 2016 08:50AM EST | DOI: 10.1002/stem.2545
Our study set out to identify the functional role of gap junctional intercellular communication (GJIC) in ES cells. The deletion of Connexin43 and Connexin45 leads to an abrogation of GJIC, which resulted in defective primitive endoderm formation in embryoid bodies (EBs). We assume that IP3 diffuses via gap junction conduits to coordinate and sustain NFATc3 activation in a synchronized manner. As a consequence of Cx43/45 deletion the IP3 mediated synchronization breaks down and the development of primitive endoderm is impaired. A similar phenotype is observed upon blocking NFAT signaling using the calcineurin inhibitor Cyclosporin A (CSA). The fluorescence pictures show Ebs on day6 of differentiation. The primitive endoderm marker Gata6 is shown in red, the pluripotency marker Nanog in green. Scale: 20 μm.